The objective of this project is the study of the structure and metabolism of several glycoprotein hormones associated with reproduction. The amino acid sequence of porcine FSH will be determined by Edman degradation of the separate reduced, carboxymethlated subunits. Certain regions of human FSH beta subunit, in dispute in the published literature, will be re-examined as well. Comparative structural analysis of the speparate carbohydrate side chains of FSH, hCG and LH will be carried out, continuing current studies to determine whether these moieties differ within subunits or continuing current studies to determine whether these moieties differ within subunits or among alpha subunits which are otherwise essentially identical. In later stages of the project, experiments will be undertaken to assign the disulfide linkages of porcine FSH. Using subunits and peptides obtained during structural analysis, region-specific immunoassays will be developed with reduced, carboxymethylated hLH subunits for use in detection and characterization of products of hormone metabolism in plasma and urine. The more sequence-specific antisera obtained in this way might be more likely to detect altered forms or fragments than the highly conformation-specific antisera against native subunits. Structure-function studies will be done to determine the effects of chemical modifications of pFSH on in vivo biological and immunological activity. Also, bioreceptor assays and adenyl cyclase assays for LH and hCG will be used to explore the use of synthetic fragments as probes for regions of the molecules involved in subunit and receptor interactions. Human pituitary extracts will be screened for free alpha subunit, and sufficient quantities isolated for chemical characterization to distinguish differences from alpha subunit found in the normal, associated state. Similarly, we will use specific hCG-beta antisera to search for and identify the hCG-like material reported to be in pituitary extracts (among a number of other non-placental sources).